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Plant and Cell Physiology Advance Access published online on July 18, 2007
Plant and Cell Physiology, doi:10.1093/pcp/pcm091
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© The Author 2007. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For Permissions, please e-mail: journals.permissions@oxfordjournals.org

An Aluminum-Activated Citrate Transporter in Barley

Jun Furukawa1, Naoki Yamaji1, Hua Wang1, Namiki Mitani1, Yoshiko Murata2, Kazuhiro Sato2, Maki Katsuhara1, Kazuyoshi Takeda1 and Jian Feng Ma1

1 Research Institute for Bioresources, Okayama University, Kurashiki 710-0046 Japan
2 Suntory Institute for Bioorganic Research, Osaka 618-8503 Japan

Corresponding author: Jian Feng MA, Research Institute for Bioresources, Okayama University, Chuo 2-20-1, Kurashiki 710-0046, Japan, Tel/Fax: +81-86-434-1209, E-mail: maj{at}rib.okayama-u.ac.jp


  Abstract

Soluble ionic Al inhibits root growth and reduces crop production on acid soils. Al-resistant cultivars of barley (Hordeum vulgare L.) detoxify Al by secreting citrate from the roots, but the responsible gene has not been identified yet. Here, we identified a gene (HvAACT1) responsible for the Al-activated citrate secretion by fine mapping combined with microarray analysis, using Al-resistant cultivar, Murasakimochi and Al-sensitive cultivar, Morex. This gene belongs to the multidrug and toxic compound extrusion (MATE) family and was constitutively expressed mainly in the roots of Al resistant barley cultivar. Heterologous expression of HvAACT1 in Xenopus oocytes showed efflux activity for 14C-labelled citrate, but not for malate. Two-electrode voltage clamp analysis also showed transport activity of citrate in the HvAACT1 expressed oocytes under Al existing condition. Over-expression of this gene in tobacco enhanced citrate secretion and Al resistance compared to the wild type plants. Transiently expressed GFP-tagged HvAACT1 was localized at the plasma membrane of the onion epidermal cells and immunostaining showed that HvAACT1 was localized at the epidermal cells in the barley root tips. A good correlation was found between the expression of HvAACT1 and citrate secretion in 10 barley cultivars differing in Al resistance. Taken together, our results demonstrate that HvAACT1 is an Al-activated citrate transporter responsible for Al resistance in barley.

Keywords: Aluminum - barley - citrate transporter - MATE - resistance - root


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